ABSTRACT: There have been few studies of the bacterial community within the bottom boundary layer (BBL) the turbulent region of the water column above the benthos in shallow seas. Typically, the BBL has large amounts of particulate organic matter suspended by turbulence, and it is often the first region of the water column to become hypoxic when oxygen declines. Communities at the surface (5 m) and in the BBL (1 to 10 m above the sea floor) were compared by terminal restriction fragment length polymorphism (T-RFLP) analysis and sequencing of the 16S rRNA gene. Multivariate statistical methods (hierarchical clustering, non-metric multidimensional scaling, and analysis of similarity (ANOSIM)) indicated that the microbial community of the BBL is distinct from the surface community. ANOSIM supported the distinction between surface and BBLs (R values 0.427 and 0.463, based on analysis with restriction enzymes BsuR1 and Hin6I, respectively, p < 0.1%). Six terminal restriction fragments showed an increase in abundance with depth. Cloning, screening and sequencing identified these as a novel environmental clade (Eastern North Pacific Chromatiales (ENPC) clade), the ARTIC96BD-19 clade of Gammaproteobacteria, the 6N14 and Agg8 clades of the phylum Planctomycetes, the OM60/NOR5 clade of Gammaproteobacteria, and uncultivated members of the Roseobacter clade in the MB11C09 and ULA23 subgroups. To the best of our knowledge, this analysis is the first to focus on the unique composition of microbial communities of the BBL in shallow, inner-shelf regions off the coast of Oregon, USA, and the first to report that an uncharacterized clade of Chromatiales is indigenous in this habitat.